RNA sequencing determines nuclear-accumulated Cav3.2iPA on transcriptional regulation of human iPSC-derived sensory neurons

By targeting the intrinsically disordered regions (IDR) of calcium channel subtype 3.2 (Cav3.2), we discovered selective T-type/Cav3.2 peptide inhibitors (Cav3.2iPAs) for AAV-mediated peripheral sensory neuron-specific analgesia. We found that Cav3.2iPAs are highly accumulated in the nuclei after expression in human induced pluripotent stem cells-derived sensory neurons (hiPSC-SNs). This study aims to determine whether nuclear-accumulated Cav3.2iPAs disturb the transcriptional regulation of hiPSC-SNs which becomes a safety concern. Overall design: RNA sequencing compares the transcriptomes in hiPSC-SNs-expressing Cav3.2iPA and hiPSC-SN-expressing Cav3.2NP (control)

本研究以钙通道亚型3.2（calcium channel subtype 3.2, Cav3.2）的内在无序区域（intrinsically disordered regions, IDR）为靶点，成功发现了可用于腺相关病毒（adeno-associated virus, AAV）介导的外周感觉神经元特异性镇痛的选择性T型/Cav3.2肽类抑制剂（Cav3.2iPAs）。
研究发现，Cav3.2iPAs在人类诱导多能干细胞衍生感觉神经元（human induced pluripotent stem cells-derived sensory neurons, hiPSC-SNs）中表达后，会高度富集于细胞核内。
本研究旨在验证富集于细胞核内的Cav3.2iPAs是否会干扰hiPSC-SNs的转录调控，这一问题已成为当前的安全性关注点。
整体实验设计：通过RNA测序，对比表达Cav3.2iPA的hiPSC-SNs与表达Cav3.2NP（对照）的hiPSC-SNs的转录组差异。